Stepwise silver-staining-based immunosorbent assay for amyloid-beta autoantibody detection

Nanomedicine (Lond). 2008 Aug;3(4):485-93. doi: 10.2217/17435889.3.4.485.


Aims: To detect amyloid-beta (Abeta) autoantibodies in a reliable and high-throughput fashion, we developed a stepwise silver-staining-based immunosorbent assay in a 96-well-plate platform.

Materials & methods: Abeta autoantibodies were incubated in an Abeta-immobilized 96-well microplate. Antihuman IgG-modified gold nanoparticle probes were then used to bind to the autoantibodies and signal enhancement was carried out with stepwise silver-staining on immobilized gold nanoparticle probes. A microplate reader was used to quantify silver-stained gold nanoparticles on a well-plate surface.

Results & discussion: Stepwise silver-staining at low temperature enables long-term silver-staining with minimal increase of background signal. This stepwise staining method helps solve the problems of one-step staining, such as nonspecific binding or nonuniformity in silver precipitation after prolonged silver-staining for signal enhancement.

Conclusion: A stepwise silver-staining strategy could be useful in minimizing nonspecific background signals. This 96-well-plate-based Abeta antibody detection assay could be useful in studying and diagnosing Alzheimer's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid beta-Peptides / immunology*
  • Autoantibodies / analysis*
  • Autoantibodies / immunology
  • Humans
  • Immunoassay / methods
  • Microscopy, Electron, Scanning
  • Nanotechnology / methods
  • Reproducibility of Results
  • Silver Staining / methods*


  • Amyloid beta-Peptides
  • Autoantibodies